Table 3.
Rapamycin distribution results in anterior segment of the rabbit eye after topical application of 0.2% rapamycin nanomicellar formulation along with the calibration curve standards (STD), and quality control (QC) standards precision & accuracy intra -day assay data, and retention time.
| Name | Nominal Conc. (ng/mL) | Retention time, min (mean, S.D.) | Cal. conc.(ng/mL) (Mean, S.D.) | (%) Accuracy | (%) Precision |
|---|---|---|---|---|---|
| CORNEA | |||||
| STD-1 | 2.3 | 3.104, 0.016 | 1.9, 0.33 | 82.6 | 17.4 |
| STD-2 | 10 | 3.017, 0.071 | 8.5, 1.37 | 85.0 | 16.1 |
| STD-3 | 100 | 3.012, 0.019 | 91.5, 7.03 | 91.6 | 7.7 |
| STD-4 | 250 | 3.018, 0.028 | 211.0, 26.0 | 85.6 | 12.7 |
| STD-5 | 500 | 3.054, 0.047 | 432.9, 62.03 | 86.6 | 14.4 |
| STD-6 | 750 | 3.030, 0.045 | 761.2, 123.0 | 101.5 | 16.2 |
| STD-7 | 850 | 3.024, 0.036 | 801.0, 124.33 | 94.6 | 15.6 |
| STD-8 | 1000 | 3.006, 0.011 | 899.9, 134.0 | 89.9 | 14.9 |
| LLOQQC | 2.3 | 3.024, 0.036 | 1.88, 0.24 | 81.6 | 12.8 |
| LQC | 10 | 3.016, 0.037 | 8.9, 1.31 | 89.0 | 14.7 |
| MQC | 480 | 3.022, 0.038 | 501.0, 71.33 | 104.3 | 14.2 |
| HQC | 800 | 3.041, 0.03 | 721.9, 81.33 | 90.1 | 7.4 |
| Results of 10 samples (mean ± S.D.): 2260.7 ± 507.1 ng/g tissue | |||||
| Iris ciliary muscle | |||||
| STD-1 | 2.3 | 3.002, 0.017 | 2.7, 0.36 | 117.4 | 13.3 |
| STD-2 | 10 | 3.000, 0.061 | 9.5, 0.90 | 95.0 | 9.5 |
| STD-3 | 100 | 3.102, 0.016 | 101.5, 5.6 | 101.5 | 5.5 |
| STD-4 | 250 | 2.995, 0.005 | 267.0, 30.0 | 106.8 | 11.2 |
| STD-5 | 500 | 3.007, 0.083 | 567.9, 32.03 | 113.6 | 5.6 |
| STD-6 | 750 | 3.028, 0.043 | 679.2, 102.0 | 90.5 | 15.0 |
| STD-7 | 850 | 3.012, 0.043 | 900.9, 125.0 | 105.6 | 13.9 |
| STD-8 | 1000 | 3.022, 0.018 | 1100.0, 124.0 | 110.0 | 12.2 |
| LLOQQC | 2.3 | 3.024, 0.036 | 2.5, 0.21 | 108.7 | 8.4 |
| LQC | 10 | 2.989, 0.061 | 10.9, 1.31 | 110.0 | 11.9 |
| MQC | 480 | 2.992, 0.034 | 550.0, 71.00 | 114.3 | 12.2 |
| HQC | 1000 | 3.001, 0.01 | 908.0, 101.33 | 113.1 | 11.4 |
| Results of 10 samples (mean ± S.D.): 585.5 ± 80.1 ng/g tissue | |||||
| Aqueous humor | |||||
| STD-1 | 2.3 | 32000, 0.002 | 1.99, 0.22 | 86.5 | 11.1 |
| STD-2 | 10 | 3.042, 0.061 | 10.9, 0.80 | 109.0 | 7.3 |
| STD-3 | 100 | 3.012, 0.019 | 111.5, 10.60 | 111.5 | 9.5 |
| STD-4 | 250 | 3.018, 0.028 | 245.0, 12.0 | 98.0 | 4.9 |
| STD-5 | 500 | 3.024, 0.047 | 570.0, 23.03 | 114.0 | 4.0 |
| STD-6 | 750 | 3.031, 0.041 | 789.2, 102.0 | 105.5 | 12.9 |
| STD-7 | 850 | 3.024, 0.037 | 906.0, 111.00 | 106.6 | 12.3 |
| STD-8 | 1000 | 3.006, 0.011 | 1113.9, 104.0 | 111.3 | 12.1 |
| LLOQQC | 2.3 | 3.004, 0.016 | 2.6, 0.32 | 89.0 | 10.1 |
| LQC | 10 | 3.018, 0.039 | 8.9, 0.90 | 89.7 | 10.1 |
| MQC | 480 | 3.032, 0.018 | 421.0, 2.60 | 87.3 | 0.6 |
| HQC | 1000 | 3.045, 0.031 | 709.9, 65.00 | 88.6 | 9.2 |
| Results of 10 samples (mean ± S.D.): below limit of quantitation (< 2.3 ng/mL) ng/mg protein | |||||
| LENS | |||||
| STD-1 | 2.3 | 2.958, 0.042 | 2.6, 0.11 | 113.0 | 4.2 |
| STD-2 | 10 | 2.958, 0.048 | 9.9, 0.89 | 99.0 | 9.0 |
| STD-3 | 100 | 2.959, 0.014 | 114.0, 8.90 | 114.0 | 7.8 |
| STD-4 | 250 | 2.952, 0.023 | 230.0, 11.0 | 92.0 | 4.7 |
| STD-5 | 500 | 2.956, 0.024 | 532.0, 20.03 | 106.2 | 3.8 |
| STD-6 | 750 | 2.979, 0.016 | 700.2, 103.0 | 93.3 | 14.7 |
| STD-7 | 850 | 2.969, 0.055 | 768.9, 102.33 | 90.6 | 13.3 |
| STD-8 | 1000 | 3.009, 0.058 | 1089.9, 112.0 | 108.9 | 10.3 |
| LLOQQC | 2.3 | 2.986, 0.065 | 2.03, 0.30 | 88.3 | 114.8 |
| LQC | 10 | 2.949, 0.036 | 9.9, 1.10 | 99.0 | 11.1 |
| MQC | 480 | 2.966, 0.019 | 521.0, 2.40 | 108.5 | 0.5 |
| HQC | 1000 | 2.984, 0.050 | 721.9, 81.33 | 113.6 | 7.8 |
| Results of 10 samples (mean ± S.D.): below limit of quantitation (<2.3 ng/mL) ng/mg protein | |||||
Ten samples of each cornea, iris ciliary muscle, lens, and aqueous humor were analyzed against each calibration curve STD and QC standards separately. BLOQ: below limit of quantitation, Low, middle and High QC: LQC, MQC and HQC, respectively.